LOCUS       sequence-243864-        8012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (RSF1010 oriR with beta-lactamase/sfGFP 
            payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8012)
  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 8012)
  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-S1
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(914..982,983..1774)
                     /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(1132..1151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1811..2524
                     /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      2584..2630
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      3405..3799
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            4141..4228
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             5467..6438
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             6949..7788
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             join(7775..8012,1..614)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
ORIGIN
        1 tgcttggccc ggaacccaag accgaaggcg gacggcagct ccggctgttc ctggaaccca
       61 agtgggaggc cgtcaccgct gatgccatgg tggtcaaagg tagctatcgg gcgctggcaa
      121 aggaaatcgg ggcagaggtc gatagtggtg gggcgctcaa gcacatacag gactgcatcg
      181 agcgcctttg gaaggtatcc atcatcgccc agaatggccg caagcggcag gggtttcggc
      241 tgctgtcgga gtacgccagc gacgaggcgg acgggcgcct gtacgtggcc ctgaacccct
      301 tgatcgcgca ggccgtcatg ggtggcggcc agcatgtgcg catcagcatg gacgaggtgc
      361 gggcgctgga cagcgaaacc gcccgcctgc tgcaccagcg gctgtgtggc tggatcgacc
      421 ccggcaaaac cggcaaggct tccatagata ccttgtgcgg ctatgtctgg ccgtcagagg
      481 ccagtggttc gaccatgcgc aagcgccgcc agcgggtgcg cgaggcgttg ccggagctgg
      541 tcgcgctggg ctggacggta accgagttcg cggcgggcaa gtacgacatc acccggccca
      601 aggcggcagg ctgacccccc ccactctatt gtaaacaaga catttttatc ttttatattc
      661 aatggcttat tttcctgcta attggtaata ccatgaaaaa taccatgctc agaaaaggct
      721 taacaatatt ttgaaaaatt gcctactgag cgctgccgca cagctccata ggccgagatg
      781 caataaatta aggagaggtt atactctaga gtagtagatt attttaggaa tttagatgtt
      841 ttgtatgaaa tagatgcttc gtatggaatt aatgaaattt ttagtcaggt aaaaaaggta
      901 ataggagaat attatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      961 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1021 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1081 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1141 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1201 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1261 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1321 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1381 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1441 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1501 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1561 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1621 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1681 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1741 tgagataggt gcctcactga ttaagcattg gtaaattata tttaaactca atttaaagta
     1801 gggagaatag atgcgtaaag gcgaagagct gttcactggt ttcgtcacta ttctggtgga
     1861 actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc
     1921 aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg
     1981 gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca
     2041 catgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac
     2101 gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga
     2161 taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct
     2221 gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca
     2281 aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca
     2341 gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga
     2401 caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca
     2461 catggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta
     2521 caaataaaaa caataggcca catgcaactg tctagaatgc gagagtaggg aactgccagg
     2581 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2641 gctcgaccag gcgtacgctt atgggtgcct ttccgcagct tggaacgcgg atggagaaga
     2701 ggagcaacgc gatctagcta tcgcggccgc gatcaagcag gtgcgacaga cgtcatacta
     2761 gatatcaagc gacttctcct atcccctggg aacacatcaa tctcaccgga gaatatcgct
     2821 ggccaaagcc ttagcgtagg attccgcccc ttcccgcaaa cgaccccaaa caggaaacgc
     2881 agctgaaacg ggaagctcaa cacccactga cgcatgggtt gttcaggcag tacttcatca
     2941 accagcaagg cggcactttc ggccatccgc cgcgccccac agctcgggca gaaaccgcga
     3001 cgcttacagc tgaaagcgac caggtgctcg gcgtggcaag actcgcagcg aacccgcaga
     3061 aagccatgct ccagccgccc gcattggaga aattcttcaa attcccgttg cacatagccc
     3121 ggcaattcct ttccctgctc tgccataagc gcagcgaatg ccgggtaata ctcgtcaacg
     3181 atctgataga gaagggtttg ctcgggtcgg tggctctggt aacgaccagt atcccgatcc
     3241 cggctggccg tcctggccgc cacatgaggc atgttccgcg tccttgcaat actgtgttta
     3301 catacagtct atcgcttagc ggaaagttct tttaccctca gccgaaatgc ctgccgttgc
     3361 tagacattgc cagccagtgc ccgtcactcc cgtactaact gtcacgaacc cctgcaataa
     3421 ctgtcacgcc cccctgcaat aactgtcacg aacccctgca ataactgtca cgcccccaaa
     3481 cctgcaaacc cagcaggggc gggggctggc ggggtgttgg aaaaatccat ccatgattat
     3541 ctaagaataa tccactaggc gcggttatca gcgcccttgt ggggcgctgc tgcccttgcc
     3601 caatatgccc ggccagaggc cggatagctg gtctattcgc tgcgctaggc tacacaccgc
     3661 cccaccgctg cgcggcaggg ggaaaggcgg gcaaagcccg ctaaacccca caccaaaccc
     3721 cgcagaaata cgctggagcg cttttagccg ctttagcggc ctttccccct acccgaaggg
     3781 tgggggcgcg tgtgcagccc cgcagggcct gtctcggtcg atcattcagc ccggctcatc
     3841 cttctggcgt ggcggcagac cgaacaaggc gcggtcgtgg tcgcgttcaa ggtacgcatc
     3901 cattgccgcc atgagccgat cctccggcca ctcgctgctg ttcaccttgg ccaaaatcat
     3961 ggcccccacc agcaccttgc gccttgtttc gttcttgcgc tcttgctgct gttcccttgc
     4021 ccgcacccgc tgaatttcgg cattgattcg cgctcgttgt tcttcgagct tggccagccg
     4081 atccgccgcc ttgttgctcc ccttaaccat cttgacaccc cattgttaat gtgctgtctc
     4141 gtaggctatc atggaggcac agcggcggca atcccgaccc tactttgtag gggagggcgc
     4201 acttaccggt ttctcttcga gaaactggcc taacggccac ccttcgggcg gtgcgctctc
     4261 cgagggccat tgcatggagc cgaaaagcaa aagcaacagc gaggcagcat ggcgatttat
     4321 caccttacgg cgaaaaccgg cagcaggtcg ggcggccaat cggccagggc caaggccgac
     4381 tacatccagc gcgaaggcaa gtatgcccgc gacatggatg aagtcttgca cgccgaatcc
     4441 gggcacatgc cggagttcgt cgagcggccc gccgactact gggatgctgc cgacctgtat
     4501 gaacgcgcca atgggcggct gttcaaggag gtcgaatttg ccctgccggt cgagctgacc
     4561 ctcgaccagc agaaggcgct ggcgtccgag ttcgcccagc acctgaccgg tgccgagcgc
     4621 ctgccgtata cgctggccat ccatgccggt ggcggcgaga acccgcactg ccacctgatg
     4681 atctccgagc ggatcaatga cggcatcgag cggcccgccg ctcagtggtt caagcggtac
     4741 aacggcaaga ccccggagaa gggcggggca cagaagaccg aagcgctcaa gcccaaggca
     4801 tggcttgagc agacccgcga ggcatgggcc gaccatgcca accgggcatt agagcgggct
     4861 ggccacgacg cccgcattga ccacagaaca cttgaggcgc agggcatcga gcgcctgccc
     4921 ggtgttcacc tggggccgaa cgtggtggag atggaaggcc ggggcatccg caccgaccgg
     4981 gcagacgtgg ccctgaacat cgacaccgcc aacgcccaga tcatcgactt acaggaatac
     5041 cgggaggcaa tagaccatga acgcaatcga cagagtgaag aaatccagag gcatcaacga
     5101 gttagcggag cagatcgaac cgctggccca gagcatggcg acactggccg acgaagcccg
     5161 gcaggtcatg agccagaccc agcaggccag cgaggcgcag gcggcggagt ggctgaaagc
     5221 ccagcgccag acaggggcgg catgggtgga gctggccaaa gagttgcggg aggtagccgc
     5281 cgaggtgagc agcgccgcgc agagcgcccg gagcgcgtcg cgggggtggc actggaagct
     5341 atggctaacc gtgatgctgg cttccatgat gcctacggtg gtgctgctga tcgcatcgtt
     5401 gctcttgctc gacctgacgc cactgacaac cgaggacggc tcgatctggc tgcgcttggt
     5461 ggcccgatga agaacgacag gactttgcag gccataggcc gacagctcaa ggccatgggc
     5521 tgtgagcgct tcgatatcgg cgtcagggac gccaccaccg gccagatgat gaaccgggaa
     5581 tggtcagccg ccgaagtgct ccagaacacg ccatggctca agcggatgaa tgcccagggc
     5641 aatgacgtgt atatcaggcc cgccgagcag gagcggcatg gtctggtgct ggtggacgac
     5701 ctcagcgagt ttgacctgga tgacatgaaa gccgagggcc gggagcctgc cctggtagtg
     5761 gaaaccagcc cgaagaacta tcaggcatgg gtcaaggtgg ccgacgccgc aggcggtgaa
     5821 cttcgggggc agattgcccg gacgctggcc agcgagtacg acgccgaccc ggccagcgcc
     5881 gacagccgcc actatggccg cttggcgggc ttcaccaacc gcaaggacaa gcacaccacc
     5941 cgcgccggtt atcagccgtg ggtgctgctg cgtgaatcca agggcaagac cgccaccgct
     6001 ggcccggcgc tggtgcagca ggctggccag cagatcgagc aggcccagcg gcagcaggag
     6061 aaggcccgca ggctggccag cctcgaactg cccgagcggc agcttagccg ccaccggcgc
     6121 acggcgctgg acgagtaccg cagcgagatg gccgggctgg tcaagcgctt cggtgatgac
     6181 ctcagcaagt gcgactttat cgccgcgcag aagctggcca gccggggccg cagtgccgag
     6241 gaaatcggca aggccatggc cgaggccagc ccagcgctgg cagagcgcaa gcccggccac
     6301 gaagcggatt acatcgagcg caccgtcagc aaggtcatgg gtctgcccag cgtccagctt
     6361 gcgcgggccg agctggcacg ggcaccggca ccccgccagc gaggcatgga caggggcggg
     6421 ccagatttca gcatgtagtg cttgcgttgg tactcacgcc tgttatacta tgagtactca
     6481 cgcacagaag ggggttttat ggaatacgaa aaaagcgctt cagggtcggt ctacctgatc
     6541 aaaagtgaca agggctattg gttgcccggt ggctttggtt atacgtcaaa caaggccgag
     6601 gctggccgct tttcagtcgc tgatatggcc agccttaacc ttgacggctg caccttgtcc
     6661 ttgttccgcg aagacaagcc tttcggcccc ggcaagtttc tcggtgactg atatgaaaga
     6721 ccaaaaggac aagcagaccg gcgacctgct ggccagccct gacgctgtac gccaagcgcg
     6781 atatgccgag cgcatgaagg ccaaagggat gcgtcagcgc aagttctggc tgaccgacga
     6841 cgaatacgag gcgctgcgcg agtgcctgga agaactcaga gcggcgcagg gcgggggtag
     6901 tgaccccgcc agcgcctaac caccaactgc ctgcaaagga ggcaatcaat ggctacccat
     6961 aagcctatca atattctgga ggcgttcgca gcagcgccgc caccgctgga ctacgttttg
     7021 cccaacatgg tggccggtac ggtcggggcg ctggtgtcgc ccggtggtgc cggtaaatcc
     7081 atgctggccc tgcaactggc cgcacagatt gcaggcgggc cggatctgct ggaggtgggc
     7141 gaactgccca ccggcccggt gatctacctg cccgccgaag acccgcccac cgccattcat
     7201 caccgcctgc acgcccttgg ggcgcacctc agcgccgagg aacggcaagc cgtggctgac
     7261 ggcctgctga tccagccgct gatcggcagc ctgcccaaca tcatggcccc ggagtggttc
     7321 gacggcctca agcgcgccgc cgagggccgc cgcctgatgg tgctggacac gctgcgccgg
     7381 ttccacatcg aggaagaaaa cgccagcggc cccatggccc aggtcatcgg tcgcatggag
     7441 gccatcgccg ccgataccgg gtgctctatc gtgttcctgc accatgccag caagggcgcg
     7501 gccatgatgg gcgcaggcga ccagcagcag gccagccggg gcagctcggt actggtcgat
     7561 aacatccgct ggcagtccta cctgtcgagc atgaccagcg ccgaggccga ggaatggggt
     7621 gtggacgacg accagcgccg gttcttcgtc cgcttcggtg tgagcaaggc caactatggc
     7681 gcaccgttcg ctgatcggtg gttcaggcgg catgacggcg gggtgctcaa gcccgccgtg
     7741 ctggagaggc agcgcaagag caagggggtg ccccgtggtg aagcctaaga acaagcacag
     7801 cctcagccac gtccggcacg acccggcgca ctgtctggcc cccggcctgt tccgtgccct
     7861 caagcggggc gagcgcaagc gcagcaagct ggacgtgacg tatgactacg gcgacggcaa
     7921 gcggatcgag ttcagcggcc cggagccgct gggcgctgat gatctgcgca tcctgcaagg
     7981 gctggtggcc atggctgggc ctaatggcct ag
//
