LOCUS       sequence-244981-        1914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes permuteposon P4 for MuA transposase-mediated circular 
            permutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1914)
  AUTHORS   Atkinson JT, Wu B, Segatori L, Silberg JJ
  TITLE     Overcoming component limitations in synthetic biology through 
            transposon-mediated protein engineering.
  JOURNAL   Methods Enzymol. 2019;621:191-212. doi: 10.1016/bs.mie.2019.02.014. 
            Epub 2019 Apr 2.
  PUBMED    31128779
REFERENCE   2  (bases 1 to 1914)
  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: "Methods 
            Enzymol."; date: "2019"; volume: "621:191-212. doi"; pages: " 
            10.1016/bs.mie.2019.02.014. Epub 2019 Apr 2"
COMMENT     Sequence Label: pMT-P4
FEATURES             Location/Qualifiers
     source          1..1914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..90)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             82..897
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALTVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(155..174)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        join(948..953,954..969,970..976)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    982..1001
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     rep_origin      complement(1280..1868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1360..1379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
       61 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
      121 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      181 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
      241 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      301 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      361 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
      421 ccagattcag gtgaaaatat tgttgatgcg ctgacagtgt tcctgcgccg gttgcattcg
      481 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
      541 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggttgg
      601 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
      661 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
      721 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
      781 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
      841 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaaaaa
      901 aaaaaacccc gcttcggcgg ggtttttttt tctcgaggat ccgaaatttg acaattaatc
      961 atcggctcgt ataatgtgtg gaattgtgag cgctcacaat taggtaccgt tttcgcattt
     1021 atcgtgaaac gctttcgcat tttgagtgag gtatatgaga tctgaagcgg cgcacgaaaa
     1081 acgcgaaagc gtttcacgat aaatgcgaaa aaaaggatct caagaagatc ctttgatttt
     1141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1201 ggggataacg caggaaagaa catgtgagca aaggccagca aaaggccagg aaccgtaaaa
     1261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1681 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1861 gatctcaaga agatcctttg atcttttcta cggggtctga cgccatggct cagt
//
