LOCUS       sequence-233939-        5340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (Himar transposon with beta-lactamase/sfGFP 
            payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  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 5340)
  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-Kh2
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..393
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            508..617
                     /note="incP origin of transfer"
     CDS             complement(938..1597)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1512..1531
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1598..1702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1770..1788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2197..2265,2266..3057)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2415..2434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3094..3807
                     /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      3867..3953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4045..4072
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
       61 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
      121 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
      181 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
      241 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
      301 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
      361 ctatcaacag gttgaactgc tgatcttcag atcctctacg ccggacgcat cgtggccgga
      421 tcaattccgt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
      481 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
      541 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
      601 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
      661 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
      721 tgaccccgaa gcagggttat gcagcggaaa acggaattga tccccctcga attgacgcgt
      781 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
      841 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
      901 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
      961 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1021 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1081 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1141 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     1201 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1261 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1321 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgta
     1381 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1441 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1501 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1561 tatcaacggt ggtatatcca gtgatttttt tctccatact cttccttttt caatattatt
     1621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1741 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcacg
     1801 cgtcctcggt acctaacagg ttggatgata agtccccggt cttcgtatgc cgtcttctgc
     1861 ttggcgcgcc ctcgagcaat tgccgaccga atttttatgt cgtaaagagg ggctttgcag
     1921 ggggtggact cagaaagatg agaatagatg actattgtag ttgaaacaca tagaaagttg
     1981 ctgatataca gaccgatacg catatcggga tgaaccatga gtacgttctt ttctcaaaaa
     2041 acataaatat tcgaaaagag atgcaataaa ttaaggagag gttatagttt taaatgatga
     2101 aaagaaatat ttagggaaga ttgtttcgac gcgaattgtt gatctggaaa atgatcacct
     2161 tatcggacaa gctttaaaat aggaggatat aaaaatatga gtattcaaca tttccgtgtc
     2221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaatt
     3061 atatttaaac tcaatttaaa gtagggagaa tagatgcgta aaggcgaaga gctgttcact
     3121 ggtttcgtca ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg
     3181 cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact
     3241 actggtaaac tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag
     3301 tgctttgctc gttatccgga ccacatgaag cagcatgact tcttcaagtc cgccatgccg
     3361 gaaggctatg tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt
     3421 gcggaagtga aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac
     3481 tttaaagaag acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat
     3541 gtttacatca ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac
     3601 aacgtggagg atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt
     3661 gatggtcctg ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa
     3721 gatccgaacg agaaacgcga tcacatggtt ctgctggagt tcgtaaccgc agcgggcatc
     3781 acgcatggta tggatgaact gtacaaataa aaacaatagg ccacatgcaa ctgtctagaa
     3841 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3901 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     3961 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     4021 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     4081 tacctgcagg gcgcgccaag cagaagacgg catacgaaga ccggggactt atcatccaac
     4141 ctgttaggat ccgttttaaa tgatgaaaag aaatatttag ggaagattgt ttcgacgcga
     4201 attgttgatc tggaaaatga tcaccttatc ggacaagctt taaaatagga ggatataaaa
     4261 atcatatgga aaaaaaggaa tttcgtgttt tgataaaata ctgttttctg aagggaaaaa
     4321 atacagtgga agcaaaaact tggcttgata atgagtttcc ggactctgcc ccagggaaat
     4381 caacaataat tgattggtat gcaaaattca agcgtggtga aatgagcacg gaggacggtg
     4441 aacgcagtgg acgcccgaaa gaggtggtta ccgacgaaaa catcaaaaaa atccacaaaa
     4501 tgattttgaa tgaccgtaaa atgaagttga tcgagatagc agaggcctta aagatatcaa
     4561 aggaacgtgt tggtcatatc attcatcaat atttggatat gcggaagctc tgtgcgaaat
     4621 gggtgccgcg cgagctcaca tttgaccaaa aacaacgacg tgttgatgat tctaagcggt
     4681 gtttgcagct gttaactcgt aatacacccg agtttttccg tcgatatgtg acaatggatg
     4741 aaacatggct ccatcactac actcctgagt ccaatcgaca gtcggctgag tggacagcga
     4801 ccggtgaacc gtctccgaag cgtggaaaga ctcaaaagtc cgctggcaaa gtaatggcct
     4861 ctgttttttg ggatgcgcat ggaataattt ttatcgatta tcttgagaag ggaaaaacca
     4921 tcaacagtga ctattatatg gcgttattgg agcgtttgaa ggtcgaaatc gcggcaaaac
     4981 ggccccacat gaagaagaaa aaagtgttgt tccaccaaga caacgcaccg tgccacaagt
     5041 cattgagaac gatggcaaaa attcatgaat tgggcttcga attgcttccc cacccgccgt
     5101 attctccaga tctggccccc agcgactttt tcttgttctc agacctcaaa aggatgctcg
     5161 cagggaaaaa atttggctgc aatgaagagg tgatcgccga aactgaggcc tattttgagg
     5221 caaaaccgaa ggagtactac caaaatggta tcaaaaaatt ggaaggtcgt tataatcgtt
     5281 gtatcgctct tgaagggaac tatgttgaat aagcggccgc caccgcggtg gaggggaatt
//
