LOCUS       sequence-496789-        7130 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  pPhlF133 driving sfGFP in pNBU2 integrative Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 7130)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM407
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..44)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      877..948
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      964..991
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1347..2060
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2067..2138
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2142..2159
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2182..2209
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4170..4961,4962..5030))
                     /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     4793..4812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6322..6710
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6881..6990
                     /note="incP origin of transfer"
ORIGIN
        1 gataaaacga aaggctcagt cgaaagactg ggcctttcgt tttacaattg ggctaccttt
       61 tttttgtttt gtttgcaatg gttaatctat tgttaaaatt taaagtttca cttgaacttt
      121 caaataatgt tcttatattt ccagtgtcga aagaaacaaa gtaggactga tcacacaaca
      181 tttaaaaaat aacattatga aagcactcga aaagagattc aaaggtaata ttaacaataa
      241 tttattttca atggcacgta ccccgagccg tagcagcatt ggtagcctgc gtagtccgca
      301 tacccataaa gcaattctga ccagcaccat tgaaatcctg aaagaatgtg gttatagcgg
      361 tctgagcatt gaaagcgttg cacgtcgtgc cggtgcaagc aaaccgacca tttatcgttg
      421 gtggaccaat aaagcagcac tgattgccga agtgtatgaa aatgaaagcg aacaggtgcg
      481 taaatttccg gatctgggta gctttaaagc cgatctggat tttctgctgc gtaatctgtg
      541 gaaagtttgg cgtgaaacca tttgtggtga agcatttcgt tgtgttattg cagaagcaca
      601 gctggaccct gcaaccctga cccagctgaa agatcagttt atggaacgtc gtcgtgagat
      661 gccgaaaaaa ctggttgaaa atgccattag caatggtgaa ctgccgaaag ataccaatcg
      721 tgaactgctg ctggatatga tttttggttt ttgttggtat cgcctgctga ccgaacagct
      781 gaccgttgaa caggatattg aagaatttac cttcctgctg attaatggtg tttgtccggg
      841 tacacagcgt taactagggc ccataccccc aggcatcaaa taaaacgaaa ggctcagtcg
      901 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
      961 actggctcac cttcgggtgg gcctttctgc gtttatacga cgtacggtgg aatctgattc
     1021 gttaccaatt gacatgatac gaaacgtacc gtatcgacgg tttatacgac gtacggtgga
     1081 atctgattcg ttaccaattg acatgatacg aaacgtaccg tatcgacggc aattgggcta
     1141 cctttttttt gttttgtttg caatggttaa tctattgtta aatgatacga aacgtaccgt
     1201 atcgttaagg tatgttctta tatttgcagt atgatacgaa acgtaccgta tcgttaaggt
     1261 gactgatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
     1321 gtaatattaa caataattta ttttcaatgc gtaaaggcga agagctgttc actggtgtcg
     1381 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     1441 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     1501 aactgccggt tccttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     1561 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     1621 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     1681 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     1741 aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     1801 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     1861 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     1921 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     1981 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     2041 gtatggatga actgtacaaa ggatctgatt acaaggacga tgatgataag gactacaaag
     2101 atgatgatga caaagattat aaggacgatg atgataaagg tcatcatcat catcatcact
     2161 aaatcctagc tgattagctc gagaaggcca tcctgacgga tggccttttt tttgtatcga
     2221 gacgactagt gtaattgcct atcttccagt gatggaacag catttgtgca ttggctgcaa
     2281 caatcagcct tacttgtgcc tgttctattt ccgaaccgac cgcttgtatg aatgcatcaa
     2341 aattcgtttt ctctacgttg gattccttgt tgctcatatt gtgatgataa tttctacaaa
     2401 tatagtcatt ggtaactatc tatgaaactg tttgatactt ttatagttga ttaaacttgt
     2461 tcatggcatt tgccttaata tcatccgcta tgtcaatgta gggtttcata gctttgtagt
     2521 cgctgtgtcc cgtccatttc atgaccacct gtgccgggat tccgagagcc agcgcattgc
     2581 agatgaatgt cctttttcct gcatgggtac tgagcaaagc gtatttgggt gtgacttcat
     2641 caatacgttc atttcccttg tagtaggttt cccgtacagg ctcgttgatt tctgccagtt
     2701 cgcccagctc tttcaggtaa tcgttcatct tctggttgct gatgacgggc agagccatgt
     2761 aattctcgaa atggatgtcc ttgtatttgt ccagtatggc tttgctgtat ttgttcagtt
     2821 caatcgtcag gctgtcggca gtcttgactg tggttatttc gatgtggtcg gacttcacat
     2881 cgcttctttt cagattgcga acatccgaat accgcaaact cgtaaagcag cagaacagga
     2941 aaacatcacg cacacgttcc aggtattgct tatccttggg tatctggtag tctttcagct
     3001 tgttcagttc atcccaagtc aggaagatta cttttttcga ggtggttttc agtttcggtt
     3061 tgaacgtatc gtatgcaatg ttctgatgat gtcctttctt gaagctccag cgcaggaacc
     3121 atttgaggaa tcccatttgc ttgccgatgg tgctgtttct catatccttg gtgtcacgca
     3181 ggaagttgac gtattcgttc aatccaaact cgttgaaata gttgaacgtt gcatcctcct
     3241 tgaactcttt gaggtggttc ctcactgctg caaatttttc ataggtggat gccgtccagt
     3301 tattctggtt accgcactct tttacaaact catcgaacac ctcccaaaag ctgacagggg
     3361 cttcttccgg ctgttcttca ctggtgtctt tcattctcat gttgaaagct tccttcaact
     3421 gttgggtcgt tggcatgacc tcctgcacct caaattcctt gaaaatattc tggatttcgg
     3481 catagtattt cagcaagtcc gtattgattt cggctgcact ttgctttagc ttgttggtac
     3541 atccgttctt tacccgctgc ttatctgcat cccatttggc tacgtcaatc cggtagcccg
     3601 ttgtaaactc gatacgttgg ctggcaaaga tgacacgcat acggatgggt acgttctcta
     3661 cgattggcac accgttcttt ttccggctct ccaatgcaaa aatgatgttg cgcttgatat
     3721 tcataattgg gtgcgtttga aattctacac ccaaatatac acccaattat tgagatagca
     3781 aaagacattt agaaacattt acttttactc tatattgtaa tttacacttg attatcagtc
     3841 gtttgcagtt ttatgatatt ctgtgaaagt ataagttcga gagcctgtct ctccgcaaaa
     3901 aacgctgaaa atcagcagat tgcaaaacaa acaccctgtt ttacacccaa gaatgtaaag
     3961 tcgggtgttt ttgttttatt taagataata caaccactac ataataaaag agtagcgata
     4021 ttaaaagaat ccgatgagaa aagactaata tttatctatc cattcagttt gatttctcag
     4081 gactttacat cgtcctgaaa gtatttgttg gtaccggtac cgaggacgcg acagctgcag
     4141 acgcgttatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5041 tttcaatcat ggccgcggga ttaaaagtcg gggattggtg aacaaaaagg tgtttctctc
     5101 tttaagagaa atatcgtttt gctaaacagt tgatattgag gtatcatttt atcgtaaaag
     5161 acatttttgc tcaacaattg cttgacggaa atcaacaaat tttagcattt tgtaaaaaag
     5221 tcgctatata atttggtgaa ttggagttat tttcatattt ttgcatcccg aagagtttct
     5281 cttaaagaga gaaacatctt ttgcatacct tttccgaccg aatttttatg tcgtaaagag
     5341 gggctttgca gggggtggac tcagaaagat gagaatagat gactattgta gttgaaacac
     5401 atagaaagtt gctgatatac agaccgatac gcatatcggg atgaaccatg agtacgttct
     5461 tttctcaaaa aacataaata ttcgaaaaga gatgcaataa attaaggaga ggttataatg
     5521 aacaaagtaa atataaaaga tagtcaaaat tttattactt caaaatatca catagaaaaa
     5581 ataatgaatt gcataagttt agatgaaaaa gataacatct ttgaaatagg tgcagggaaa
     5641 ggtcatttta ctgctggatt ggtaaagaga tgtaattttg taacggcgat agaaattgat
     5701 tctaaattat gtgaggtaac tcgtaataag ctcttaaatt atcctaacta tcaaatagta
     5761 aatgatgata tactgaaatt tacatttcct agccacaatc catataaaat atttggcagc
     5821 ataccttaca acataagcac aaatataatt cgaaaaattg tttttgaaag ttcagccaca
     5881 ataagttatt taatagtgga atatggtttt gctaaaatgt tattagatac aaacagatca
     5941 ctagcattgc tgttaatggc agaggtagat atttctatat tagcaaaaat tcctaggtat
     6001 tatttccatc caaaacctaa agtggatagc acattaattg tattaaaaag aaagccagca
     6061 aaaatggcat ttaaagagag aaaaaaatat gaaacttttg taatgaaatg ggttaacaaa
     6121 gagtacgaaa aactgtttac aaaaaatcaa tttaataaag ctttaaaaca tgcgagaata
     6181 tatgatataa acaatattag tttcgaacaa tttgtatcgc tatttaatag ttataaaata
     6241 tttaacggct aaaaacaata ggccacatgc aactgtaaat gtttacgcgg gtaccgacac
     6301 cgcggtggag gggaattccc atgtcagccg ttaagtgttc ctgtgtcact caaaattgct
     6361 ttgagaggct ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt
     6421 aaaccttaaa agctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct
     6481 tagaggctat ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt
     6541 acgtgaaaca tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg
     6601 tttagttcgt taaacatgag agcttagtac gttaaacatg agagcttagt acgtgaaaca
     6661 tgagagctta gtacgtacta tcaacaggtt gaactgctga tcttcagatc ctctacgccg
     6721 gacgcatcgt ggccggatca attccgtttt ccgctgcata accctgcttc ggggtcatta
     6781 tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg
     6841 ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag
     6901 gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca
     6961 acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat gagggcaagc
     7021 ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcacg gaattgatcc
     7081 ccctcgaatt ggaattcgtt cctcgattca gatctaaagg atccaatcta
//
