LOCUS       sequence-241990-        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (pBBR1 oriR with beta-lactamase/sfGFP payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  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 6236)
  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-B2
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     48..64
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     49..65
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             333..1043
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGFVTILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDEL*K"
     primer_bind     complement(1068..1090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1784..2446)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2447..3218
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             complement(join(4721..5512,5513..5581))
                     /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     5344..5363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5582..5686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5754..5772
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5810..5832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    6102..6123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6138..6143,6144..6161,6162..6168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6176..6192
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6181..6203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6200..6216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6200..6216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     join(6235..6236,1..19)
                     /label=T3
                     /note="T3 promoter, forward primer"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct gggtaccggg ccccccctcg aggtcgacgg
       61 tatcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta
      121 ctccgtcaag ccgtcactca ttggatccgg gagagcttca acggcgcttc tacccatttg
      181 cttggaaagg atgaggagca ggaagaaatt ccgtccccaa tgcgacggcc ctttacatcc
      241 atgttgtttg atagtataat ggatacggat tgaccaaatt gttcatttag tcagtttgaa
      301 ggatgaggag tatgctatct tcggtgctgc agcgtaaagg cgaagagctg ttcactggtt
      361 tcgtcactat tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg
      421 gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg
      481 gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct
      541 ttgctcgtta tccggaccac atgaagcagc atgacttctt caagtccgcc atgccggaag
      601 gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg
      661 aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta
      721 aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt
      781 acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg
      841 tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg
      901 gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc
      961 cgaacgagaa acgcgatcac atggttctgc tggagttcgt aaccgcagcg ggcatcacgc
     1021 atggtatgga tgaactgtaa aaataacgcg aattccagca cggccgtcgt tttacaacgt
     1081 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     1141 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1201 ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1261 gttaaatcag ctcatttttt aaccaatagg ccgtactgcg atgagtggca gggcggggcg
     1321 taattttttt aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt
     1381 gataataagc ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc gtcggttcag
     1441 ggcagggtcg ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg
     1501 gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag gctctccccg
     1561 tggaggtaat aattgacgat atgatcattt attctgcctc ccagagcctg ataaaaacgg
     1621 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     1681 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgaggcggc tacagccgat
     1741 agtctggaac agcgcactta cgggttgctg cgcaacccaa gtgctaccgg cgcggcagcg
     1801 tgacccgtgt cggcggctcc aacggctcgc catcgtccag aaaacacggc tcatcgggca
     1861 tcggcaggcg ctgctgcccg cgccgttccc attcctccgt ttcggtcaag gctggcaggt
     1921 ctggttccat gcccggaatg ccgggctggc tgggcggctc ctcgccgggg ccggtcggta
     1981 gttgctgctc gcccggatac agggtcggga tgcggcgcag gtcgccatgc cccaacagcg
     2041 attcgtcctg gtcgtcgtga tcaaccacca cggcggcact gaacaccgac aggcgcaact
     2101 ggtcgcgggg ctggccccac gccacgcggt cattgaccac gtaggccgac acggtgccgg
     2161 ggccgttgag cttcacgacg gagatccagc gctcggccac caagtccttg actgcgtatt
     2221 ggaccgtccg caaagaacgt ccgatgagct tggaaagtgt cttctggctg accaccacgg
     2281 cgttctggtg gcccatctgc gccacgaggt gatgcagcag cattgccgcc gtgggtttcc
     2341 tcgcaataag cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc cagtgaccgg
     2401 gcttgttctt ggcttgaatg ccgatttctc tggactgcgt ggccatgctt atctccatgc
     2461 ggtaggggtg ccgcacggtt gcggcaccat gcgcaatcag ctgcaacttt tcggcagcgc
     2521 gacaacaatt atgcgttgcg taaaagtggc agtcaattac agattttctt taacctacgc
     2581 aatgagctat tgcggggggt gccgcaatga gctgttgcgt accccccttt tttaagttgt
     2641 tgatttttaa gtctttcgca tttcgcccta tatctagttc tttggtgccc aaagaagggc
     2701 acccctgcgg ggttccccca cgccttcggc gcggctcccc ctccggcaaa aagtggcccc
     2761 tccggggctt gttgatcgac tgcgcggcct tcggccttgc ccaaggtggc gctgccccct
     2821 tggaaccccc gcactcgccg ccgtgaggct cggggggcag gcgggcgggc ttcgcccttc
     2881 gactgccccc actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg
     2941 cggtcgctgc gcgagccttg acccgccttc cacttggtgt ccaaccggca agcgaagcgc
     3001 gcaggccgca ggccggaggc ttttccccag agaaaattaa aaaaattgat ggggcaaggc
     3061 cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca
     3121 atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt
     3181 gtccacgggc cgagcgaagc gagccagccg gtggccgctc gcggccatcg tccacatatc
     3241 cacgggctgg caagggagcg cagcgaccgc gcagggcgaa gcccggagag caagcccgta
     3301 gggcgccgca gccgccgtag gcggtcacga ctttgcgaag caaagtctag tgagtatact
     3361 caagcattga gtggcccgcc ggaggcaccg ccttgcgctg cccccgtcga gccggttgga
     3421 caccaaaagg gaggggcagg catggcggca tacgcgatca tgcgatgcaa gaagctggcg
     3481 aaaatgggca acgtggcggc cagtctcaag cacgcctacc gcgagcgcga gacgcccaac
     3541 gctgacgcca gcaggacgcc agagaacgag cactgggcgg ccagcagcac cgatgaagcg
     3601 atgggccgac tgcgcgagtt gctgccagag aagcggcgca aggacgctgt gttggcggtc
     3661 gagtacgtca tgacggccag cccggaatgg tggaagtcgg ccagccaaga acagcaggcg
     3721 gcgttcttcg agaaggcgca caagtggctg gcggacaagt acggggcgga tcgcatcgtg
     3781 acggccagca tccaccgtga cgaaaccagc ccgcacatga ccgcgttcgt ggtgccgctg
     3841 acgcaggacg gcaggctgtc ggccaaggag ttcatcggca acaaagcgca gatgacccgc
     3901 gaccagacca cgtttgcggc cgctgtggcc gatctagggc tgcaacgggg catcgagggc
     3961 agcaaggcac gtcacacgcg cattcaggcg ttctacgagg ccctggagcg gccaccagtg
     4021 ggccacgtca ccatcagccc gcaagcggtc gagccacgcg cctatgcacc gcagggattg
     4081 gccgaaaagc tgggaatctc aaagcgcgtt gagacgccgg aagccgtggc cgaccggctg
     4141 acaaaagcgg ttcggcaggg gtatgagcct gccctacagg ccgccgcagg agcgcgtgag
     4201 atgcgcaaga aggccgatca agcccaagag acggcccgag accttcggga gcgcctgaag
     4261 cccgttctgg acgccctggg gccgttgaat cgggatatgc aggccaaggc cgccgcgatc
     4321 atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg aagtccagcg ccagaaacag
     4381 gcccagcgcc agcaggaacg cgggcgcgca catttccccg aaaagtgcca cctgcggcgt
     4441 tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt
     4501 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
     4561 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
     4621 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
     4681 cggtgctctc cggagactgc gagatcatag atatagatct ttaccaatgc ttaatcagtg
     4741 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4801 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4861 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4921 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4981 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5041 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5101 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagttcc ttcggtcctc
     5161 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5221 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5281 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5341 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5401 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5461 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5521 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5581 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5641 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5701 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5761 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5821 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc catgggcaaa
     5881 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     5941 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagttttt atgcatgcgc
     6001 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6061 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgc
//
