LOCUS       sequence-241988-        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-B1
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     60..80
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        62..80
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     109..125
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     110..126
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             394..1104
                     /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(1129..1151)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1845..2507)
                     /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      2508..3279
                     /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(4782..5573,5574..5642))
                     /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     5405..5424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5643..5747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5815..5833
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5871..5893)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    6163..6184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6199..6204,6205..6222,6223..6229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
       61 caattaaccc tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg
      121 gtatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
      181 actccgtcaa gccgtcactc attggatccg attgcattag gttttagttt cttgtataat
      241 gcttaatgtt ggtcactgac aggctacgat acggaaggtt gctcacgccc ggcccctttg
      301 ccatggctag tgtgtggaaa tttccgagga gcaagtctat ttccaaaaat gggcgaaaaa
      361 ggaggtaata caatgatgct accgactctg cagcgtaaag gcgaagagct gttcactggt
      421 ttcgtcacta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
      481 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
      541 ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
      601 tttgctcgtt atccggacca catgaagcag catgacttct tcaagtccgc catgccggaa
      661 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg
      721 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt
      781 aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
      841 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac
      901 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
      961 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     1021 ccgaacgaga aacgcgatca catggttctg ctggagttcg taaccgcagc gggcatcacg
     1081 catggtatgg atgaactgta aaaataacgc gaattccagc acggccgtcg ttttacaacg
     1141 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1201 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1261 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1321 tgttaaatca gctcattttt taaccaatag gccgtactgc gatgagtggc agggcggggc
     1381 gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg cctgaataag
     1441 tgataataag cggatgaatg gcagaaattc gaaagcaaat tcgacccggt cgtcggttca
     1501 gggcagggtc gttaaatagc cgcttatgtc tattgctggt ttaccggttt attgactacc
     1561 ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc agtttgctca ggctctcccc
     1621 gtggaggtaa taattgacga tatgatcatt tattctgcct cccagagcct gataaaaacg
     1681 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     1741 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgaggcgg ctacagccga
     1801 tagtctggaa cagcgcactt acgggttgct gcgcaaccca agtgctaccg gcgcggcagc
     1861 gtgacccgtg tcggcggctc caacggctcg ccatcgtcca gaaaacacgg ctcatcgggc
     1921 atcggcaggc gctgctgccc gcgccgttcc cattcctccg tttcggtcaa ggctggcagg
     1981 tctggttcca tgcccggaat gccgggctgg ctgggcggct cctcgccggg gccggtcggt
     2041 agttgctgct cgcccggata cagggtcggg atgcggcgca ggtcgccatg ccccaacagc
     2101 gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac tgaacaccga caggcgcaac
     2161 tggtcgcggg gctggcccca cgccacgcgg tcattgacca cgtaggccga cacggtgccg
     2221 gggccgttga gcttcacgac ggagatccag cgctcggcca ccaagtcctt gactgcgtat
     2281 tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg tcttctggct gaccaccacg
     2341 gcgttctggt ggcccatctg cgccacgagg tgatgcagca gcattgccgc cgtgggtttc
     2401 ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt ccgtttgcac ccagtgaccg
     2461 ggcttgttct tggcttgaat gccgatttct ctggactgcg tggccatgct tatctccatg
     2521 cggtaggggt gccgcacggt tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg
     2581 cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta cagattttct ttaacctacg
     2641 caatgagcta ttgcgggggg tgccgcaatg agctgttgcg tacccccctt ttttaagttg
     2701 ttgattttta agtctttcgc atttcgccct atatctagtt ctttggtgcc caaagaaggg
     2761 cacccctgcg gggttccccc acgccttcgg cgcggctccc cctccggcaa aaagtggccc
     2821 ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc
     2881 ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg cttcgccctt
     2941 cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc aagccgctgc
     3001 gcggtcgctg cgcgagcctt gacccgcctt ccacttggtg tccaaccggc aagcgaagcg
     3061 cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga tggggcaagg
     3121 ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta gccggtgggc
     3181 aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt ccagcagggt
     3241 tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc gtccacatat
     3301 ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga gcaagcccgt
     3361 agggcgccgc agccgccgta ggcggtcacg actttgcgaa gcaaagtcta gtgagtatac
     3421 tcaagcattg agtggcccgc cggaggcacc gccttgcgct gcccccgtcg agccggttgg
     3481 acaccaaaag ggaggggcag gcatggcggc atacgcgatc atgcgatgca agaagctggc
     3541 gaaaatgggc aacgtggcgg ccagtctcaa gcacgcctac cgcgagcgcg agacgcccaa
     3601 cgctgacgcc agcaggacgc cagagaacga gcactgggcg gccagcagca ccgatgaagc
     3661 gatgggccga ctgcgcgagt tgctgccaga gaagcggcgc aaggacgctg tgttggcggt
     3721 cgagtacgtc atgacggcca gcccggaatg gtggaagtcg gccagccaag aacagcaggc
     3781 ggcgttcttc gagaaggcgc acaagtggct ggcggacaag tacggggcgg atcgcatcgt
     3841 gacggccagc atccaccgtg acgaaaccag cccgcacatg accgcgttcg tggtgccgct
     3901 gacgcaggac ggcaggctgt cggccaagga gttcatcggc aacaaagcgc agatgacccg
     3961 cgaccagacc acgtttgcgg ccgctgtggc cgatctaggg ctgcaacggg gcatcgaggg
     4021 cagcaaggca cgtcacacgc gcattcaggc gttctacgag gccctggagc ggccaccagt
     4081 gggccacgtc accatcagcc cgcaagcggt cgagccacgc gcctatgcac cgcagggatt
     4141 ggccgaaaag ctgggaatct caaagcgcgt tgagacgccg gaagccgtgg ccgaccggct
     4201 gacaaaagcg gttcggcagg ggtatgagcc tgccctacag gccgccgcag gagcgcgtga
     4261 gatgcgcaag aaggccgatc aagcccaaga gacggcccga gaccttcggg agcgcctgaa
     4321 gcccgttctg gacgccctgg ggccgttgaa tcgggatatg caggccaagg ccgccgcgat
     4381 catcaaggcc gtgggcgaaa agctgctgac ggaacagcgg gaagtccagc gccagaaaca
     4441 ggcccagcgc cagcaggaac gcgggcgcgc acatttcccc gaaaagtgcc acctgcggcg
     4501 ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg
     4561 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
     4621 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
     4681 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
     4741 ccggtgctct ccggagactg cgagatcata gatatagatc tttaccaatg cttaatcagt
     4801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5101 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5161 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagttc cttcggtcct
     5221 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5281 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5341 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5401 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5461 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5521 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5581 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5641 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5701 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5761 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5821 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5881 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag ccatgggcaa
     5941 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     6001 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtttt tatgcatgcg
     6061 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6121 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6181 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaa
//
