LOCUS       sequence-241991-        6238 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 6238)
  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 6238)
  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-B3
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     98..118
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        100..118
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     147..163
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     148..164
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             434..1144
                     /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(1169..1191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1885..2547)
                     /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      2548..3319
                     /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(4822..5613,5614..5682))
                     /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     5445..5464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5683..5787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5855..5873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5911..5933)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    6203..6224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      121 acaaaagctg ggtaccgggc cccccctcga ggtcgacggt atcgatgcat aatgtgcctg
      181 tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcactcat
      241 tggatccgtg aaggatacgg ctgcggcact tcgacatcgc cccatgtggc ggctttgaac
      301 tgggcttatg aaacgcgttc acaacctttt ttgaccatcg gcgcgaacgt ggtatcatgc
      361 gttcagcttt tgcccataca tactacgtgc tcaatctagg aggatttcat acatgcgtga
      421 tcattttctg cagcgtaaag gcgaagagct gttcactggt ttcgtcacta ttctggtgga
      481 actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc
      541 aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg
      601 gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca
      661 catgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac
      721 gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga
      781 taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct
      841 gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca
      901 aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg gcagcgtgca
      961 gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga
     1021 caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca
     1081 catggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta
     1141 aaaataacgc gaattccagc acggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1201 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1261 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat
     1321 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1381 taaccaatag gccgtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     1441 attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg
     1501 gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc
     1561 cgcttatgtc tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg
     1621 cttctcaaat gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga
     1681 tatgatcatt tattctgcct cccagagcct gataaaaacg gtgaatccgt tagcgaggtg
     1741 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     1801 aggcagacaa ggtatagggc ggcgaggcgg ctacagccga tagtctggaa cagcgcactt
     1861 acgggttgct gcgcaaccca agtgctaccg gcgcggcagc gtgacccgtg tcggcggctc
     1921 caacggctcg ccatcgtcca gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc
     1981 gcgccgttcc cattcctccg tttcggtcaa ggctggcagg tctggttcca tgcccggaat
     2041 gccgggctgg ctgggcggct cctcgccggg gccggtcggt agttgctgct cgcccggata
     2101 cagggtcggg atgcggcgca ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg
     2161 atcaaccacc acggcggcac tgaacaccga caggcgcaac tggtcgcggg gctggcccca
     2221 cgccacgcgg tcattgacca cgtaggccga cacggtgccg gggccgttga gcttcacgac
     2281 ggagatccag cgctcggcca ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg
     2341 tccgatgagc ttggaaagtg tcttctggct gaccaccacg gcgttctggt ggcccatctg
     2401 cgccacgagg tgatgcagca gcattgccgc cgtgggtttc ctcgcaataa gcccggccca
     2461 cgcctcatgc gctttgcgtt ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat
     2521 gccgatttct ctggactgcg tggccatgct tatctccatg cggtaggggt gccgcacggt
     2581 tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc
     2641 gtaaaagtgg cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg
     2701 tgccgcaatg agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc
     2761 atttcgccct atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc
     2821 acgccttcgg cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga
     2881 ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc
     2941 gccgtgaggc tcggggggca ggcgggcggg cttcgccctt cgactgcccc cactcgcata
     3001 ggcttgggtc gttccaggcg cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt
     3061 gacccgcctt ccacttggtg tccaaccggc aagcgaagcg cgcaggccgc aggccggagg
     3121 cttttcccca gagaaaatta aaaaaattga tggggcaagg ccgcaggccg cgcagttgga
     3181 gccggtgggt atgtggtcga aggctgggta gccggtgggc aatccctgtg gtcaagctcg
     3241 tgggcaggcg cagcctgtcc atcagcttgt ccagcagggt tgtccacggg ccgagcgaag
     3301 cgagccagcc ggtggccgct cgcggccatc gtccacatat ccacgggctg gcaagggagc
     3361 gcagcgaccg cgcagggcga agcccggaga gcaagcccgt agggcgccgc agccgccgta
     3421 ggcggtcacg actttgcgaa gcaaagtcta gtgagtatac tcaagcattg agtggcccgc
     3481 cggaggcacc gccttgcgct gcccccgtcg agccggttgg acaccaaaag ggaggggcag
     3541 gcatggcggc atacgcgatc atgcgatgca agaagctggc gaaaatgggc aacgtggcgg
     3601 ccagtctcaa gcacgcctac cgcgagcgcg agacgcccaa cgctgacgcc agcaggacgc
     3661 cagagaacga gcactgggcg gccagcagca ccgatgaagc gatgggccga ctgcgcgagt
     3721 tgctgccaga gaagcggcgc aaggacgctg tgttggcggt cgagtacgtc atgacggcca
     3781 gcccggaatg gtggaagtcg gccagccaag aacagcaggc ggcgttcttc gagaaggcgc
     3841 acaagtggct ggcggacaag tacggggcgg atcgcatcgt gacggccagc atccaccgtg
     3901 acgaaaccag cccgcacatg accgcgttcg tggtgccgct gacgcaggac ggcaggctgt
     3961 cggccaagga gttcatcggc aacaaagcgc agatgacccg cgaccagacc acgtttgcgg
     4021 ccgctgtggc cgatctaggg ctgcaacggg gcatcgaggg cagcaaggca cgtcacacgc
     4081 gcattcaggc gttctacgag gccctggagc ggccaccagt gggccacgtc accatcagcc
     4141 cgcaagcggt cgagccacgc gcctatgcac cgcagggatt ggccgaaaag ctgggaatct
     4201 caaagcgcgt tgagacgccg gaagccgtgg ccgaccggct gacaaaagcg gttcggcagg
     4261 ggtatgagcc tgccctacag gccgccgcag gagcgcgtga gatgcgcaag aaggccgatc
     4321 aagcccaaga gacggcccga gaccttcggg agcgcctgaa gcccgttctg gacgccctgg
     4381 ggccgttgaa tcgggatatg caggccaagg ccgccgcgat catcaaggcc gtgggcgaaa
     4441 agctgctgac ggaacagcgg gaagtccagc gccagaaaca ggcccagcgc cagcaggaac
     4501 gcgggcgcgc acatttcccc gaaaagtgcc acctgcggcg ttgtgacaat ttaccgaaca
     4561 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     4621 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     4681 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     4741 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctct ccggagactg
     4801 cgagatcata gatatagatc tttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5221 ccccatgttg tgcaaaaaag cggttagttc cttcggtcct ccgatcgttg tcagaagtaa
     5281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5881 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5941 cacagcttgt ctgtaagcgg atgccgggag ccatgggcaa atattatacg caaggcgaca
     6001 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     6061 gcagaatgct taatgaatta caacagtttt tatgcatgcg cccaatacgc aaaccgcctc
     6121 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6181 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggc
//
