LOCUS       sequence-243869-        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-S2
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(118..186,187..978)
                     /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(336..355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1015..1728
                     /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      1788..1834
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      2609..3003
                     /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            3345..3432
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4671..5642
                     /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             6153..6992
                     /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             6979..7830
                     /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 ggttatagtt ttaaatgatg aaaagaaata tttagggaag attgtttcga cgcgaattgt
       61 tgatctggaa aatgatcacc ttatcggaca agctttaaaa taggaggata taaaaatatg
      121 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      181 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      241 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      301 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      361 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      421 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      481 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      541 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      601 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      661 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      721 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      781 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      841 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      901 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      961 ctgattaagc attggtaaat tatatttaaa ctcaatttaa agtagggaga atagatgcgt
     1021 aaaggcgaag agctgttcac tggtttcgtc actattctgg tggaactgga tggtgatgtc
     1081 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
     1141 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
     1201 acgctgactt atggtgttca gtgctttgct cgttatccgg accacatgaa gcagcatgac
     1261 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
     1321 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
     1381 attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa
     1441 tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa
     1501 gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac
     1561 cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc
     1621 acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcacatggt tctgctggag
     1681 ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaata aaaacaatag
     1741 gccacatgca actgtctaga atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     1801 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttggctcga ccaggcgtac
     1861 gcttatgggt gcctttccgc agcttggaac gcggatggag aagaggagca acgcgatcta
     1921 gctatcgcgg ccgcgatcaa gcaggtgcga cagacgtcat actagatatc aagcgacttc
     1981 tcctatcccc tgggaacaca tcaatctcac cggagaatat cgctggccaa agccttagcg
     2041 taggattccg ccccttcccg caaacgaccc caaacaggaa acgcagctga aacgggaagc
     2101 tcaacaccca ctgacgcatg ggttgttcag gcagtacttc atcaaccagc aaggcggcac
     2161 tttcggccat ccgccgcgcc ccacagctcg ggcagaaacc gcgacgctta cagctgaaag
     2221 cgaccaggtg ctcggcgtgg caagactcgc agcgaacccg tagaaagcca tgctccagcc
     2281 gcccgcattg gagaaattct tcaaattccc gttgcacata gcccggcaat tcctttccct
     2341 gctctgccat aagcgcagcg aatgccgggt aatactcgtc aacgatctga tagagaaggg
     2401 tttgctcggg tcggtggctc tggtaacgac cagtatcccg atcccggctg gccgtcctgg
     2461 ccgccacatg aggcatgttc cgcgtccttg caatactgtg tttacataca gtctatcgct
     2521 tagcggaaag ttcttttacc ctcagccgaa atgcctgccg ttgctagaca ttgccagcca
     2581 gtgcccgtca ctcccgtact aactgtcacg aacccctgca ataactgtca cgcccccctg
     2641 caataactgt cacgaacccc tgcaataact gtcacgcccc caaacctgca aacccagcag
     2701 gggcgggggc tggcggggtg ttggaaaaat ccatccatga ttatctaaga ataatccact
     2761 aggcgcggtt atcagcgccc ttgtggggcg ctgctgccct tgcccaatat gcccggccag
     2821 aggccggata gctggtctat tcgctgcgct aggctacaca ccgccccacc gctgcgcggc
     2881 agggggaaag gcgggcaaag cccgctaaac cccacaccaa accccgcaga aatacgctgg
     2941 agcgctttta gccgctttag cggcctttcc ccctacccga agggtggggg cgcgtgtgca
     3001 gccccgcagg gcctgtctcg gtcgatcatt cagcccggct catccttctg gcgtggcggc
     3061 agaccgaaca aggcgcggtc gtggtcgcgt tcaaggtacg catccattgc cgccatgagc
     3121 cgatcctccg gccactcgct gctgttcacc ttggccaaaa tcatggcccc caccagcacc
     3181 ttgcgccttg tttcgttctt gcgctcttgc tgctgttccc ttgcccgcac ccgctgaatt
     3241 tcggcattga ttcgcgctcg ttgttcttcg agcttggcca gccgatccgc cgccttgttg
     3301 ctccccttaa ccatcttgac accccattgt taatgtgctg tctcgtaggc tatcatggag
     3361 gcacagcggc ggcaatcccg accctacttt gtaggggagg gcgcacttac cggtttctct
     3421 tcgagaaact ggcctaacgg ccacccttcg ggcggtgcgc tctccgaggg ccattgcatg
     3481 gagccgaaaa gcaaaagcaa cagcgaggca gcatggcgat ttatcacctt acggcgaaaa
     3541 ccggcagcag gtcgggcggc caatcggcca gggccaaggc cgactacatc cagcgcgaag
     3601 gcaagtatgc ccgcgacatg gatgaagtct tgcacgccga atccgggcac atgccggagt
     3661 tcgtcgagcg gcccgccgac tactgggatg ctgccgacct gtatgaacgc gccaatgggc
     3721 ggctgttcaa ggaggtcgaa tttgccctgc cggtcgagct gaccctcgac cagcagaagg
     3781 cgctggcgtc cgagttcgcc cagcacctga ccggtgccga gcgcctgccg tatacgctgg
     3841 ccatccatgc cggtggcggc gagaacccgc actgccacct gatgatctcc gagcggatca
     3901 atgacggcat cgagcggccc gccgctcagt ggttcaagcg gtacaacggc aagaccccgg
     3961 agaagggcgg ggcacagaag accgaagcgc tcaagcccaa ggcatggctt gagcagaccc
     4021 gcgaggcatg ggccgaccat gccaaccggg cattagagcg ggctggccac gacgcccgca
     4081 ttgaccacag aacacttgag gcgcagggca tcgagcgcct gcccggtgtt cacctggggc
     4141 cgaacgtggt ggagatggaa ggccggggca tccgcaccga ccgggcagac gtggccctga
     4201 acatcgacac cgccaacgcc cagatcatcg acttacagga ataccgggag gcaatagacc
     4261 atgaacgcaa tcgacagagt gaagaaatcc agaggcatca acgagttagc ggagcagatc
     4321 gaaccgctgg cccagagcat ggcgacactg gccgacgaag cccggcaggt catgagccag
     4381 acccagcagg ccagcgaggc gcaggcggcg gagtggctga aagcccagcg ccagacaggg
     4441 gcggcatggg tggagctggc caaagagttg cgggaggtag ccgccgaggt gagcagcgcc
     4501 gcgcagagcg cccggagcgc gtcgcggggg tggcactgga agctatggct aaccgtgatg
     4561 ctggcttcca tgatgcctac ggtggtgctg ctgatcgcat cgttgctctt gctcgacctg
     4621 acgccactga caaccgagga cggctcgatc tggctgcgct tggtggcccg atgaagaacg
     4681 acaggacttt gcaggccata ggccgacagc tcaaggccat gggctgtgag cgcttcgata
     4741 tcggcgtcag ggacgccacc accggccaga tgatgaaccg ggaatggtca gccgccgaag
     4801 tgctccagaa cacgccatgg ctcaagcgga tgaatgccca gggcaatgac gtgtatatca
     4861 ggcccgccga gcaggagcgg catggtctgg tgctggtgga cgacctcagc gagtttgacc
     4921 tggatgacat gaaagccgag ggccgggagc ctgccctggt agtggaaacc agcccgaaga
     4981 actatcaggc atgggtcaag gtggccgacg ccgcaggcgg tgaacttcgg gggcagattg
     5041 cccggacgct ggccagcgag tacgacgccg acccggccag cgccgacagc cgccactatg
     5101 gccgcttggc gggcttcacc aaccgcaagg acaagcacac cacccgcgcc ggttatcagc
     5161 cgtgggtgct gctgcgtgaa tccaagggca agaccgccac cgctggcccg gcgctggtgc
     5221 agcaggctgg ccagcagatc gagcaggccc agcggcagca ggagaaggcc cgcaggctgg
     5281 ccagcctcga actgcccgag cggcagctta gccgccaccg gcgcacggcg ctggacgagt
     5341 accgcagcga gatggccggg ctggtcaagc gcttcggtga tgacctcagc aagtgcgact
     5401 ttatcgccgc gcagaagctg gccagccggg gccgcagtgc cgaggaaatc ggcaaggcca
     5461 tggccgaggc cagcccagcg ctggcagagc gcaagcccgg ccacgaagcg gattacatcg
     5521 agcgcaccgt cagcaaggtc atgggtctgc ccagcgtcca gcttgcgcgg gccgagctgg
     5581 cacgggcacc ggcaccccgc cagcgaggca tggacagggg cgggccagat ttcagcatgt
     5641 agtgcttgcg ttggtactca cgcctgttat actatgagta ctcacgcaca gaagggggtt
     5701 ttatggaata cgaaaaaagc gcttcagggt cggtctacct gatcaaaagt gacaagggct
     5761 attggttgcc cggtggcttt ggttatacgt caaacaaggc cgaggctggc cgcttttcag
     5821 tcgctgatat ggccagcctt aaccttgacg gctgcacctt gtccttgttc cgcgaagaca
     5881 agcctttcgg ccccggcaag tttctcggtg actgatatga aagaccaaaa ggacaagcag
     5941 accggcgacc tgctggccag ccctgacgct gtacgccaag cgcgatatgc cgagcgcatg
     6001 aaggccaaag ggatgcgtca gcgcaagttc tggctgaccg acgacgaata cgaggcgctg
     6061 cgcgagtgcc tggaagaact cagagcggcg cagggcgggg gtagtgaccc cgccagcgcc
     6121 taaccaccaa ctgcctgcaa aggaggcaat caatggctac ccataagcct atcaatattc
     6181 tggaggcgtt cgcagcagcg ccgccaccgc tggactacgt tttgcccaac atggtggccg
     6241 gtacggtcgg ggcgctggtg tcgcccggtg gtgccggtaa atccatgctg gccctgcaac
     6301 tggccgcaca gattgcaggc gggccggatc tgctggaggt gggcgaactg cccaccggcc
     6361 cggtgatcta cctgcccgcc gaagacccgc ccaccgccat tcatcaccgc ctgcatgccc
     6421 ttggggcgca cctcagcgcc gaggaacggc aagccgtggc tgacggcctg ctgatccagc
     6481 cgctgatcgg cagcctgccc aacatcatgg ccccggagtg gttcgacggc ctcaagcgcg
     6541 ccgccgaggg ccgccgcctg atggtgctgg acacgctgcg ccggttccac atcgaggaag
     6601 aaaacgccag cggccccatg gcccaggtca tcggtcgcat ggaggccatc gccgccgata
     6661 ccgggtgctc tatcgtgttc ctgcaccatg ccagcaaggg cgcggccatg atgggcgcag
     6721 gcgaccagca gcaggccagc cggggcagct cggtactggt cgataacatc cgctggcagt
     6781 cctacctgtc gagcatgacc agcgccgagg ccgaggaatg gggtgtggac gacgaccagc
     6841 gccggttctt cgtccgcttc ggtgtgagca aggccaacta tggcgcaccg ttcgctgatc
     6901 ggtggttcag gcggcatgac ggcggggtgc tcaagcccgc cgtgctggag aggcagcgca
     6961 agagcaaggg ggtgccccgt ggtgaagcct aagaacaagc acagcctcag ccacgtccgg
     7021 cacgacccgg cgcactgtct ggcccccggc ctgttccgtg ccctcaagcg gggcgagcgc
     7081 aagcgcagca agctggacgt gacgtatgac tacggcgacg gcaagcggat cgagttcagc
     7141 ggcccggagc cgctgggcgc tgatgatctg cgcatcctgc aagggctggt ggccatggct
     7201 gggcctaatg gcctagtgct tggcccggaa cccaagaccg aaggcggacg gcagctccgg
     7261 ctgttcctgg aacccaagtg ggaggccgtc accgctgatg ccatggtggt caaaggtagc
     7321 tatcgggcgc tggcaaagga aatcggggca gaggtcgata gtggtggggc gctcaagcac
     7381 atacaggact gcatcgagcg cctttggaag gtatccatca tcgcccagaa tggccgcaag
     7441 cggcaggggt ttcggctgct gtcggagtac gccagcgacg aggcggacgg gcgcctgtac
     7501 gtggccctga accccttgat cgcgcaggcc gtcatgggtg gcggccagca tgtgcgcatc
     7561 agcatggacg aggtgcgggc gctggacagc gaaaccgccc gcctgctgca ccagcggctg
     7621 tgtggctgga tcgaccccgg caaaaccggc aaggcttcca tagatacctt gtgcggctat
     7681 gtctggccgt cagaggccag tggttcgacc atgcgcaagc gccgccagcg ggtgcgcgag
     7741 gcgttgccgg agctggtcgc gctgggctgg acggtaaccg agttcgcggc gggcaagtac
     7801 gacatcaccc ggcccaaggc ggcaggctga ccccccccac tctattgtaa acaagacatt
     7861 tttatctttt atattcaatg gcttattttc ctgctaattg gtaataccat gaaaaatacc
     7921 atgctcagaa aaggcttaac aatattttga aaaattgcct actgagcgct gccgcacagc
     7981 tccataggcc gagatgcaat aaattaagga ga
//
