LOCUS       sequence-234113-        5749 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 5749)
  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 5749)
  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-Ah2
FEATURES             Location/Qualifiers
     source          1..5749
                     /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(1347..2006)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1921..1940
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2007..2111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2179..2197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(2606..2674,2675..3466)
                     /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(2824..2843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3503..4216
                     /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      4276..4362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4454..4481
                     /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 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     1081 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     1141 cagggttatg cagcggaaaa cggaattgat ccccctcgaa ttgacgcgta accagcaata
     1201 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc
     1261 gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa
     1321 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     1381 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1441 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1501 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1561 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1621 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1681 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1741 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgtaa ttccggatga
     1801 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1861 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1921 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1981 gtatatccag tgattttttt ctccatactc ttcctttttc aatattattg aagcatttat
     2041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2161 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcacgc gtcctcggta
     2221 cctaacaggt tggatgataa gtccccggtc ttcgtatgcc gtcttctgct tggcgcgccc
     2281 tcgagcaatt gccgaccgaa tttttatgtc gtaaagaggg gctttgcagg gggtggactc
     2341 agaaagatga gaatagatga ctattgtagt tgaaacacat agaaagttgc tgatatacag
     2401 accgatacgc atatcgggat gaaccatgag tacgttcttt tctcaaaaaa cataaatatt
     2461 cgaaaagaga tgcaataaat taaggagagg ttatagtttt aaatgatgaa aagaaatatt
     2521 tagggaagat tgtttcgacg cgaattgttg atctggaaaa tgatcacctt atcggacaag
     2581 ctttaaaata ggaggatata aaaatatgag tattcaacat ttccgtgtcg cccttattcc
     2641 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2701 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2761 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2821 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2881 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2941 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3001 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3061 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3121 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3181 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3241 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3301 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3361 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3421 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaatta tatttaaact
     3481 caatttaaag tagggagaat agatgcgtaa aggcgaagag ctgttcactg gtttcgtcac
     3541 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
     3601 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
     3661 gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
     3721 ttatccggac cacatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
     3781 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
     3841 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga
     3901 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
     3961 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
     4021 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
     4081 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
     4141 gaaacgcgat cacatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat
     4201 ggatgaactg tacaaataaa aacaataggc cacatgcaac tgtctagaat gcgagagtag
     4261 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     4321 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     4381 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     4441 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acctgcaggg
     4501 cgcgccaagc agaagacggc atacgaagac cggggactta tcatccaacc tgttaggatc
     4561 cgttttaaat gatgaaaaga aatatttagg gaagattgtt tcgacgcgaa ttgttgatct
     4621 ggaaaatgat caccttatcg gacaagcttt aaaataggag gatataaaaa tcatatggaa
     4681 aaaaaggaat ttcgtgtttt gataaaatac tgttttctga agggaaaaaa tacagtggaa
     4741 gcaaaaactt ggcttgataa tgagtttccg gactctgccc cagggaaatc aacaataatt
     4801 gattggtatg caaaattcaa gcgtggtgaa atgagcacgg aggacggtga acgcagtgga
     4861 cgcccgaaag aggtggttac cgacgaaaac atcaaaaaaa tccacaaaat gattttgaat
     4921 gaccgtaaaa tgaagttgat cgagatagca gaggccttaa agatatcaaa ggaacgtgtt
     4981 ggtcatatca ttcatcaata tttggatatg cggaagctct gtgcgaaatg ggtgccgcgc
     5041 gagctcacat ttgaccaaaa acaacgacgt gttgatgatt ctaagcggtg tttgcagctg
     5101 ttaactcgta atacacccga gtttttccgt cgatatgtga caatggatga aacatggctc
     5161 catcactaca ctcctgagtc caatcgacag tcggctgagt ggacagcgac cggtgaaccg
     5221 tctccgaagc gtggaaagac tcaaaagtcc gctggcaaag taatggcctc tgttttttgg
     5281 gatgcgcatg gaataatttt tatcgattat cttgagaagg gaaaaaccat caacagtgac
     5341 tattatatgg cgttattgga gcgtttgaag gtcgaaatcg cggcaaaacg gccccacatg
     5401 aagaagaaaa aagtgttgtt ccaccaagac aacgcaccgt gccacaagtc attgagaacg
     5461 atggcaaaaa ttcatgaatt gggcttcgaa ttgcttcccc acccgccgta ttctccagat
     5521 ctggccccca gcgacttttt cttgttctca gacctcaaaa ggatgctcgc agggaaaaaa
     5581 tttggctgca atgaagaggt gatcgccgaa actgaggcct attttgaggc aaaaccgaag
     5641 gagtactacc aaaatggtat caaaaaattg gaaggtcgtt ataatcgttg tatcgctctt
     5701 gaagggaact atgttgaata agcggccgcc accgcggtgg aggggaatt
//
