LOCUS       sequence-234116-        5561 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     rep_origin      130..675
                     /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     576..595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     792..809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            945..1054
                     /note="incP origin of transfer"
     CDS             complement(1375..2184)
                     /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     2098..2117
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2185..2289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2357..2375
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             3343..4056
                     /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      4116..4202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4294..4321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcggccgcca ccgcggtgga ggggaattaa caacttatat cgtatggggc tgacttcagg
       61 tgctacattt gaagagataa attgcactga aatctagaaa tattttatct gattaataag
      121 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
      181 cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
      241 ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
      301 tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
      361 catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
      421 gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
      481 tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
      541 gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt
      601 cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg
      661 cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc
      721 tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa
      781 cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatccagatc ctctacgccg
      841 gacgcatcgt ggccggatca attccgtgat aggtgggctg cccttcctgg ttggcttggt
      901 ttcatcagcc atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag
      961 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
     1021 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
     1081 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
     1141 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaacg gaattgatcc
     1201 ccctcgaatt gacgcgtaac cagcaataga cataagcggc tatttaacga ccctgccctg
     1261 aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact
     1321 tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattagaa
     1381 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     1441 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     1501 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     1561 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     1621 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     1681 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     1741 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     1801 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     1861 taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg
     1921 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     1981 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     2041 tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc
     2101 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     2161 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     2221 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2281 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     2341 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcacgcgt cctcggtacc
     2401 taacaggttg gatgataagt ccccggtctt cgtatgccgt cttctgcttg gcgcgccctc
     2461 gagcaattgc cgaccgaatt tttatgtcgt aaagaggggc tttgcagggg gtggactcag
     2521 aaagatgaga atagatgact attgtagttg aaacacgcaa gtgttcaaga agttattaag
     2581 tcgggagtgc agtcgaagtg ggcaagttga aaaattcaca aaaatgtggt ataatatctt
     2641 tgttcattag agcgataaac ttgaatttga gagggaactt agatggtatt tgaaaaaatt
     2701 gataaaaata gttggaacag aaaagagtat tttgaccact actttgcaag tgtaccttgt
     2761 acatacagca tgaccgttaa agtggatatc acacaaataa aggaaaaggg aatgaaacta
     2821 tatcctgcaa tgctttatta tattgcaatg attgtaaacc gccattcaga gtttaggacg
     2881 gcaatcaatc aagatggtga attggggata tatgatgaga tgataccaag ctatacaata
     2941 tttcacaatg atactgaaac attttccagc ctttggactg agtgtaagtc tgactttaaa
     3001 tcatttttag cagattatga aagtgatacg caacggtatg gaaacaatca tagaatggaa
     3061 ggaaagccaa atgctccgga aaacattttt aatgtatcta tgataccgtg gtcaaccttc
     3121 gatggcttta atctgaattt gcagaaagga tatgattatt tgattcctat ttttactatg
     3181 gggaaatatt ataaagaaga taacaaaatt atacttcctt tggcaattca agttcatcac
     3241 gcagtatgtg acggatttca catttgccgt tttgtaaacg aattgcagga attgataaat
     3301 agttaaatta tatttaaact caatttaaag tagggagaat agatgcgtaa aggcgaagag
     3361 ctgttcactg gtttcgtcac tattctggtg gaactggatg gtgatgtcaa cggtcataag
     3421 ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc
     3481 atctgtacta ctggtaaact gccggtacct tggccgactc tggtaacgac gctgacttat
     3541 ggtgttcagt gctttgctcg ttatccggac cacatgaagc agcatgactt cttcaagtcc
     3601 gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac
     3661 aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa
     3721 ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata caattttaac
     3781 agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa
     3841 attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact
     3901 ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt
     3961 ctgtctaaag atccgaacga gaaacgcgat cacatggttc tgctggagtt cgtaaccgca
     4021 gcgggcatca cgcatggtat ggatgaactg tacaaataaa aacaataggc cacatgcaac
     4081 tgtctagaat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4141 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4201 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4261 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4321 ttgcgtttct acctgcaggg cgcgccaagc agaagacggc atacgaagac cggggactta
     4381 tcatccaacc tgttaggatc cctctagagt agtagattat tttaggaatt tagatgtttt
     4441 gtatgaaata gatgcttcgt atggaattaa tgaaattttt agtcaggtaa aaaaggtaat
     4501 aggagaatat tcatatggaa aaaaaggaat ttcgtgtttt gataaaatac tgttttctga
     4561 agggaaaaaa tacagtggaa gcaaaaactt ggcttgataa tgagtttccg gactctgccc
     4621 cagggaaatc aacaataatt gattggtatg caaaattcaa gcgtggtgaa atgagcacgg
     4681 aggacggtga acgcagtgga cgcccgaaag aggtggttac cgacgaaaac atcaaaaaaa
     4741 tccacaaaat gattttgaat gaccgtaaaa tgaagttgat cgagatagca gaggccttaa
     4801 agatatcaaa ggaacgtgtt ggtcatatca ttcatcaata tttggatatg cggaagctct
     4861 gtgcgaaatg ggtgccgcgc gagctcacat ttgaccaaaa acaacgacgt gttgatgatt
     4921 ctaagcggtg tttgcagctg ttaactcgta atacacccga gtttttccgt cgatatgtga
     4981 caatggatga aacatggctc catcactaca ctcctgagtc caatcgacag tcggctgagt
     5041 ggacagcgac cggtgaaccg tctccgaagc gtggaaagac tcaaaagtcc gctggcaaag
     5101 taatggcctc tgttttttgg gatgcgcatg gaataatttt tatcgattat cttgagaagg
     5161 gaaaaaccat caacagtgac tattatatgg cgttattgga gcgtttgaag gtcgaaatcg
     5221 cggcaaaacg gccccacatg aagaagaaaa aagtgttgtt ccaccaagac aacgcaccgt
     5281 gccacaagtc attgagaacg atggcaaaaa ttcatgaatt gggcttcgaa ttgcttcccc
     5341 acccgccgta ttctccagat ctggccccca gcgacttttt cttgttctca gacctcaaaa
     5401 ggatgctcgc agggaaaaaa tttggctgca atgaagaggt gatcgccgaa actgaggcct
     5461 attttgaggc aaaaccgaag gagtactacc aaaatggtat caaaaaattg gaaggtcgtt
     5521 ataatcgttg tatcgctctt gaagggaact atgttgaata a
//
