LOCUS       sequence-243854-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (Himar transposon with catP/sfGFP payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  AUTHORS   Ronda C, Chen SP, Cabral V, Yaung SJ, Wang HH
  TITLE     Metagenomic engineering of the mammalian gut microbiome in situ.
  JOURNAL   Nat Methods. 2019 Feb;16(2):167-170. doi: 10.1038/s41592-018-0301-y.
            Epub 2019 Jan 14.
  PUBMED    30643213
REFERENCE   2  (bases 1 to 5561)
  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"; doi: 
            "10.1038/s41592-018-0301-y"; journalName: "Nat Methods"; date: 
            "2019-02"; volume: "16"; issue: "2"; pages: "167-170"
COMMENT     Sequence Label: pGT-Ah5
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             895..1608
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGFVTILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1668..1754
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1846..1873
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      3243..3788
                     /direction=RIGHT
                     /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     3689..3708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3905..3922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            4058..4167
                     /note="incP origin of transfer"
     CDS             complement(4488..5297)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5211..5230
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5298..5402)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5470..5488
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tggcgcgccc tcgagcaatt gccgaccgaa tttttatgtc gtaaagaggg gctttgcagg
       61 gggtggactc agaaagatga gaatagatga ctattgtagt tgaaacacgc aagtgttcaa
      121 gaagttatta agtcgggagt gcagtcgaag tgggcaagtt gaaaaattca caaaaatgtg
      181 gtataatatc tttgttcatt agagcgataa acttgaattt gagagggaac ttagatggta
      241 tttgaaaaaa ttgataaaaa tagttggaac agaaaagagt attttgacca ctactttgca
      301 agtgtacctt gtacatacag catgaccgtt aaagtggata tcacacaaat aaaggaaaag
      361 ggaatgaaac tatatcctgc aatgctttat tatattgcaa tgattgtaaa ccgccattca
      421 gagtttagga cggcaatcaa tcaagatggt gaattgggga tatatgatga gatgatacca
      481 agctatacaa tatttcacaa tgatactgaa acattttcca gcctttggac tgagtgtaag
      541 tctgacttta aatcattttt agcagattat gaaagtgata cgcaacggta tggaaacaat
      601 catagaatgg aaggaaagcc aaatgctccg gaaaacattt ttaatgtatc tatgataccg
      661 tggtcaacct tcgatggctt taatctgaat ttgcagaaag gatatgatta tttgattcct
      721 atttttacta tggggaaata ttataaagaa gataacaaaa ttatacttcc tttggcaatt
      781 caagttcatc acgcagtatg tgacggattt cacatttgcc gttttgtaaa cgaattgcag
      841 gaattgataa atagttaaat tatatttaaa ctcaatttaa agtagggaga atagatgcgt
      901 aaaggcgaag agctgttcac tggtttcgtc actattctgg tggaactgga tggtgatgtc
      961 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
     1021 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
     1081 acgctgactt atggtgttca gtgctttgct cgttatccgg accacatgaa gcagcatgac
     1141 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
     1201 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
     1261 attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa
     1321 tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa
     1381 gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac
     1441 cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc
     1501 acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcacatggt tctgctggag
     1561 ttcgtaaccg cagcgggcat cacacatggt atggatgaac tgtacaaata aaaacaatag
     1621 gccacatgca actgtctaga atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     1681 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1741 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     1801 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     1861 cggatggcct ttttgcgttt ctacctgcag ggcgcgccaa gcagaagacg gcatacgaag
     1921 accggggact tatcatccaa cctgttagga tccctctaga gtagtagatt attttaggaa
     1981 tttagatgtt ttgtatgaaa tagatgcttc gtatggaatt aatgaaattt ttagtcaggt
     2041 aaaaaaggta ataggagaat attcatatgg aaaaaaagga atttcgtgtt ttgataaaat
     2101 actgttttct gaagggaaaa aatacagtgg aagcaaaaac ttggcttgat aatgagtttc
     2161 cggactctgc cccagggaaa tcaacaataa ttgattggta tgcaaaattc aagcgtggtg
     2221 aaatgagcac ggaggacggt gaacgcagtg gacgcccgaa agaggtggtt accgacgaaa
     2281 acatcaaaaa aatccacaaa atgattttga atgaccgtaa aatgaagttg atcgagatag
     2341 cagaggcctt aaagatatca aaggaacgtg ttggtcatat cattcatcaa tatttggata
     2401 tgcggaagct ctgtgcgaaa tgggtgccgc gcgagctcac atttgaccaa aaacaacgac
     2461 gtgttgatga ttctaagcgg tgtttgcagc tgttaactcg taatacaccc gagtttttcc
     2521 gtcgatatgt gacaatggat gaaacatggc tccatcacta cactcctgag tccaatcgac
     2581 agtcggctga gtggacagcg accggtgaac cgtctccgaa gcgtggaaag actcaaaagt
     2641 ccgctggcaa agtaatggcc tctgtttttt gggatgcgca tggaataatt tttatcgatt
     2701 atcttgagaa gggaaaaacc atcaacagtg actattatat ggcgttattg gagcgtttga
     2761 aggtcgaaat cgcggcaaaa cggccccaca tgaagaagaa aaaagtgttg ttccaccaag
     2821 acaacgcacc gtgccacaag tcattgagaa cgatggcaaa aattcatgaa ttgggcttcg
     2881 aattgcttcc ccacccgccg tattctccag atctggcccc cagcgacttt ttcttgttct
     2941 cagacctcaa aaggatgctc gcagggaaaa aatttggctg caatgaagag gtgatcgccg
     3001 aaactgaggc ctattttgag gcaaaaccga aggagtacta ccaaaatggt atcaaaaaat
     3061 tggaaggtcg ttataatcgt tgtatcgctc ttgaagggaa ctatgttgaa taagcggccg
     3121 ccaccgcggt ggaggggaat taacaactta tatcgtatgg ggctgacttc aggtgctaca
     3181 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     3241 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3301 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     3361 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     3421 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     3481 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3541 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     3601 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     3661 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     3721 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     3781 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     3841 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     3901 gcgtagcgag tcagtgagcg aggaagcgga atatatccag atcctctacg ccggacgcat
     3961 cgtggccgga tcaattccgt gataggtggg ctgcccttcc tggttggctt ggtttcatca
     4021 gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga
     4081 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     4141 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     4201 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     4261 atcgctataa tgaccccgaa gcagggttat gcagcggaaa acggaattga tccccctcga
     4321 attgacgcgt atccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     4381 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     4441 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta gaaaaactca
     4501 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     4561 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     4621 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     4681 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     4741 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     4801 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     4861 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     4921 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     4981 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     5041 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     5101 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     5161 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     5221 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     5281 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     5341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5401 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5461 acctataaaa ataggcgtat cacgaggccc tttcgtcacg cgtcctcggt acctaacagg
     5521 ttggatgata agtccccggt cttcgtatgc cgtcttctgc t
//
