LOCUS       sequence-237513-        3451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  engineered Mu transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3451)
  AUTHORS   Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA, 
            Savage DF
  TITLE     Profiling of engineering hotspots identifies an allosteric 
            CRISPR-Cas9 switch.
  JOURNAL   Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528.
  PUBMED    27136077
REFERENCE   2  (bases 1 to 3451)
  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: "Nat 
            Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528."
COMMENT     Sequence Label: pBLO 4.01
FEATURES             Location/Qualifiers
     source          1..3451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1616..1687
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1703..1730
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1758..1775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1892..2437)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1972..1991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2551..2645)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2666..3325)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3240..3259
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3326..3428)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 taggatccgt tttcgcattt atcgtgaaac gctttcgcat tttgagtgaa ttatatgaag
      781 cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg
      841 gtagcttatt cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa
      901 tccgttaaag agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg
      961 attgactttt tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta
     1021 cctaaatata gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga
     1081 gaattacaaa aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta
     1141 gctagtcatt atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt
     1201 gtggagcagc ataagcatta tttagatgag attattgagc aaatcagtga attttctaag
     1261 cgtgttattt tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga
     1321 gacaaaccaa tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt
     1381 ggagctcccg ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct
     1441 acaaaagaag ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca
     1501 cgcattgatt tgagtcagct aggaggtgac aagcttgatt gattgaacga aaaacgcgaa
     1561 agcgtttcac gataaatgcg aaaacggatc ctctcgagta aggatctcca ggcatcaaat
     1621 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1681 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta
     1741 gggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     1801 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     1861 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     1921 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     1981 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     2041 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     2101 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     2161 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     2221 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     2281 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     2341 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     2401 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     2461 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     2521 catacgatat aagttgttac tagtgcttgg attctcacca ataaaaaacg cccggcggca
     2581 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     2641 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     2701 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     2761 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     2821 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     2881 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     2941 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3001 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3061 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     3121 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     3181 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     3241 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     3301 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     3361 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     3421 gccgatcaac gtctcatttt cgccagatat c
//
