LOCUS       sequence-241979-        5753 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 5753)
  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 5753)
  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-Ah1
FEATURES             Location/Qualifiers
     source          1..5753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..647
                     /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     548..567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     764..781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            917..1026
                     /note="incP origin of transfer"
     CDS             complement(1351..2010)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1925..1944
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2011..2115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2183..2201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2610..2678,2679..3470)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDTGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMDADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2828..2847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3507..4220
                     /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      4280..4366
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4458..4485
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
       61 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
      121 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
      181 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
      241 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
      301 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
      361 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
      421 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
      481 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
      541 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
      601 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
      661 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
      721 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
      781 ggaagcggaa tatatccaga tcctctacgc cggacgcatc gtggccggat caattccgtg
      841 ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct
      901 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
      961 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     1021 ctgcccgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
     1081 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
     1141 gaagcagggt tatgcagcgg aaaacggaat tgatccccct cgaattgacg cgtaaccagc
     1201 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc
     1261 tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt
     1321 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
     1381 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     1441 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     1501 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     1561 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     1621 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     1681 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     1741 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gtaattccgg
     1801 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     1861 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     1921 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     1981 ggtggtatat ccagtgattt ttttctccat actcttcctt tttcaatatt attgaagcat
     2041 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2101 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2161 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc acgcgtcctc
     2221 ggtacctaac aggttggatg ataagtcccc ggtcttcgta tgccgtcttc tgcttggcgc
     2281 gccctcgagc aattgccgac cgaattttta tgtcgtaaag aggggctttg cagggggtgg
     2341 actcagaaag atgagaatag atgactattg tagttgaaac acatagaaag ttgctgatat
     2401 acagaccgat acgcatatcg ggatgaacca tgagtacgtt cttttctcaa aaaacataaa
     2461 tattcgaaaa gagatgcaat aaattaagga gaggttatac tctagagtag tagattattt
     2521 taggaattta gatgttttgt atgaaataga tgcttcgtat ggaattaatg aaatttttag
     2581 tcaggtaaaa aaggtaatag gagaatatta tgagtattca acatttccgt gtcgccctta
     2641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2701 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2761 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2821 aagttctgct atgtggcgcg gtattatccc gtattgacac cgggcaagag caactcggtc
     2881 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3001 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3181 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggatg
     3241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa attatattta
     3481 aactcaattt aaagtaggga gaatagatgc gtaaaggcga agagctgttc actggtttcg
     3541 tcactattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     3601 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     3661 aactgccggt accttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     3721 ctcgttatcc ggaccacatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     3781 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     3841 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     3901 aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     3961 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     4021 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     4081 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     4141 acgagaaacg cgatcacatg gttctgctgg agttcgtaac cgcagcgggc atcacacatg
     4201 gtatggatga actgtacaaa taaaaacaat aggccacatg caactgtcta gaatgcgaga
     4261 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4321 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     4381 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     4441 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacctgc
     4501 agggcgcgcc aagcagaaga cggcatacga agaccgggga cttatcatcc aacctgttag
     4561 gatccctcta gagtagtaga ttattttagg aatttagatg ttttgtatga aatagatgct
     4621 tcgtatggaa ttaatgaaat ttttagtcag gtaaaaaagg taataggaga atattcatat
     4681 ggaaaaaaag gaatttcgtg ttttgataaa atactgtttt ctgaagggaa aaaatacagt
     4741 ggaagcaaaa acttggcttg ataatgagtt tccggactct gccccaggga aatcaacaat
     4801 aattgattgg tatgcaaaat tcaagcgtgg tgaaatgagc acggaggacg gtgaacgcag
     4861 tggacgcccg aaagaggtgg ttaccgacga aaacatcaaa aaaatccaca aaatgatttt
     4921 gaatgaccgt aaaatgaagt tgatcgagat agcagaggcc ttaaagatat caaaggaacg
     4981 tgttggtcat atcattcatc aatatttgga tatgcggaag ctctgtgcga aatgggtgcc
     5041 gcgcgagctc acatttgacc aaaaacaacg acgtgttgat gattctaagc ggtgtttgca
     5101 gctgttaact cgtaatacac ccgagttttt ccgtcgatat gtgacaatgg atgaaacatg
     5161 gctccatcac tacactcctg agtccaatcg acagtcggct gagtggacag cgaccggtga
     5221 accgtctccg aagcgtggaa agactcaaaa gtccgctggc aaagtaatgg cctctgtttt
     5281 ttgggatgcg catggaataa tttttatcga ttatcttgag aagggaaaaa ccatcaacag
     5341 tgactattat atggcgttat tggagcgttt gaaggtcgaa atcgcggcaa aacggcccca
     5401 catgaagaag aaaaaagtgt tgttccacca agacaacgca ccgtgccaca agtcattgag
     5461 aacgatggca aaaattcatg aattgggctt cgaattgctt ccccacccgc cgtattctcc
     5521 agatctggcc cccagcgact ttttcttgtt ctcagacctc aaaaggatgc tcgcagggaa
     5581 aaaatttggc tgcaatgaag aggtgatcgc cgaaactgag gcctattttg aggcaaaacc
     5641 gaaggagtac taccaaaatg gtatcaaaaa attggaaggt cgttataatc gttgtatcgc
     5701 tcttgaaggg aactatgttg aataagcggc cgccaccgcg gtggagggga att
//
