LOCUS       sequence-245938-        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"
     CDS             join(1..69,70..861)
                     /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(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             898..1611
                     /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      1671..1757
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1849..1876
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      3149..3537
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            3652..3761
                     /note="incP origin of transfer"
     CDS             complement(4082..4741)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4656..4675
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4742..4846)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4914..4932
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta aattatattt aaactcaatt taaagtaggg agaatagatg
      901 cgtaaaggcg aagagctgtt cactggtttc gtcactattc tggtggaact ggatggtgat
      961 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
     1021 ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc gactctggta
     1081 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccacat gaagcagcat
     1141 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
     1201 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
     1261 cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg
     1321 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
     1381 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
     1441 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
     1501 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcacat ggttctgctg
     1561 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa ataaaaacaa
     1621 taggccacat gcaactgtct agaatgcgag agtagggaac tgccaggcat caaataaaac
     1681 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1741 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     1801 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     1861 tgacggatgg cctttttgcg tttctacctg cagggcgcgc caagcagaag acggcatacg
     1921 aagaccgggg acttatcatc caacctgtta ggatccgttt taaatgatga aaagaaatat
     1981 ttagggaaga ttgtttcgac gcgaattgtt gatctggaaa atgatcacct tatcggacaa
     2041 gctttaaaat aggaggatat aaaaatcata tggaaaaaaa ggaatttcgt gttttgataa
     2101 aatactgttt tctgaaggga aaaaatacag tggaagcaaa aacttggctt gataatgagt
     2161 ttccggactc tgccccaggg aaatcaacaa taattgattg gtatgcaaaa ttcaagcgtg
     2221 gtgaaatgag cacggaggac ggtgaacgca gtggacgccc gaaagaggtg gttaccgacg
     2281 aaaacatcaa aaaaatccac aaaatgattt tgaatgaccg taaaatgaag ttgatcgaga
     2341 tagcagaggc cttaaagata tcaaaggaac gtgttggtca tatcattcat caatatttgg
     2401 atatgcggaa gctctgtgcg aaatgggtgc cgcgcgagct cacatttgac caaaaacaac
     2461 gacgtgttga tgattctaag cggtgtttgc agctgttaac tcgtaataca cccgagtttt
     2521 tccgtcgata tgtgacaatg gatgaaacat ggctccatca ctacactcct gagtccaatc
     2581 gacagtcggc tgagtggaca gcgaccggtg aaccgtctcc gaagcgtgga aagactcaaa
     2641 agtccgctgg caaagtaatg gcctctgttt tttgggatgc gcatggaata atttttatcg
     2701 attatcttga gaagggaaaa accatcaaca gtgactatta tatggcgtta ttggagcgtt
     2761 tgaaggtcga aatcgcggca aaacggcccc acatgaagaa gaaaaaagtg ttgttccacc
     2821 aagacaacgc accgtgccac aagtcattga gaacgatggc aaaaattcat gaattgggct
     2881 tcgaattgct tccccacccg ccgtattctc cagatctggc ccccagcgac tttttcttgt
     2941 tctcagacct caaaaggatg ctcgcaggga aaaaatttgg ctgcaatgaa gaggtgatcg
     3001 ccgaaactga ggcctatttt gaggcaaaac cgaaggagta ctaccaaaat ggtatcaaaa
     3061 aattggaagg tcgttataat cgttgtatcg ctcttgaagg gaactatgtt gaataagcgg
     3121 ccgccaccgc ggtggagggg aattcccatg tcagccgtta agtgttcctg tgtcactcaa
     3181 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
     3241 aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt
     3301 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
     3361 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
     3421 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
     3481 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc
     3541 tacgccggac gcatcgtggc cggatcaatt ccgtgatagg tgggctgccc ttcctggttg
     3601 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
     3661 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     3721 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     3781 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     3841 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaacggaa
     3901 ttgatccccc tcgaattgac gcgtaaccag caatagacat aagcggctat ttaacgaccc
     3961 tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc atccgcttat
     4021 tatcacttat tcaggcgtag caaccaggcg tttaagggca ccaataactg ccttaaaaaa
     4081 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     4141 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     4201 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     4261 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     4321 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     4381 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     4441 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     4501 caccgtcttt cattgccata cgtaattccg gatgagcatt catcaggcgg gcaagaatgt
     4561 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     4621 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     4681 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     4741 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4801 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4861 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4921 gtatcacgag gccctttcgt cacgcgtcct cggtacctaa caggttggat gataagtccc
     4981 cggtcttcgt atgccgtctt ctgcttggcg cgccctcgag caattgccga ccgaattttt
     5041 atgtcgtaaa gaggggcttt gcagggggtg gactcagaaa gatgagaata gatgactatt
     5101 gtagttgaaa cacatagaaa gttgctgata tacagaccga tacgcatatc gggatgaacc
     5161 atgagtacgt tcttttctca aaaaacataa atattcgaaa agagatgcaa taaattaagg
     5221 agaggttata gttttaaatg atgaaaagaa atatttaggg aagattgttt cgacgcgaat
     5281 tgttgatctg gaaaatgatc accttatcgg acaagcttta aaataggagg atataaaaat
//
